Human liver cancer high-transfer cell strain with stable expression of fluorescent protein and construction method thereof

Abstract

The invention belongs to the field of micro-organism animal cell line and relates to a human hepatoma cell line which can emit high-intensity red or green fluorescence and has high transferring ability of lung and lymph node metastasis, and a method for establishing the same. The method comprises the following steps: using the human hepatoma cell line HCCLM3 and HCCLM6 which have high transferring ability of the lung and lymph node metastasis as mother cells, performing cotransfection on plasmid DNA of 239 cells through slow virus packaging plasmids to obtain false slow virus particles by expressing red or green fluorescent protein genes through eucaryon, and infecting liver cancer cell strains of the mother cells to obtain the chromosome integrated hepatoma cell line which has high transferring ability of the lung and lymph node metastasis and can stably expressing the red or the green fluorescence. The human hepatoma cell line which has high transferring ability of the lung and lymph node metastasis in vitro can be applied to the tracer studies on tumor cells, the molecular mechanism studies on the recurrence and transferring of liver cancer, as well as the pre-clinical drug efficacy studies on new anti-tumor drugs, thus the human hepatoma cell line has wide application prospect.

Description

technical field [0001] The invention belongs to the field of microbial animal cell lines, and relates to a fluorescently labeled human liver cancer cell line with high metastases, in particular to human liver cancer cells capable of stably expressing red fluorescent protein (RFP) or green fluorescent protein (GFP) and having high lung and lymph node metastatic ability Strains, and methods for their establishment. Background technique [0002] Primary hepatocellular carcinoma is one of the most common malignant tumors in my country and even in the world. According to statistics, about 200,000 people die of liver cancer every year in my country, accounting for about 50% of the global liver cancer deaths. Primary hepatocellular carcinoma has increasingly become a major malignant disease that seriously affects people's health. In recent years, governments, relevant medical and scientific research institutions, and drug R&D and production companies have increased their clinical...

AI Technical Summary

Problems solved by technology

Second, traditional therapies cannot completely eliminate liver cancer cells and have relatively large toxic and side effects, which limits their repeated clinical application

Third, at present, there is still a lack of long-term stable expression in vivo and in vitro, high experimental repeatability, fluorescent protein-labeled liver cancer cell lines, and high lymph node and lung metastasis at home and abroad, so as to meet the early indication of the biological behavior of liver cancer recurrence and metastasis. Fluorescent labeling research model for tracking research, molecular mechanism research and rapid screening of new anti-liver cancer drugs

The following major defects generally exist in this technology: 1) in the in vitro cell culture system, the fluorescent protein coding gene is not easy to transfect tumor cells in the G0-G1 phase, which accounts for about 70% of the total number of cells, so the transfection efficiency of the target gene is low; 2 ) Tumor cells transfected with fluorescent protein-encoding genes often require a continuous and complex in vitro screening process of antibiotics such as neomycin (G418), which can easily cause cell contamination; 3) integration of fluorescent protein-encoding genes on tumor cell chromosomes The efficiency is very low, and the gene encoding the fluorescent protein is easily lost during continuous cell passage in vitro, so it is not easy to obtain long-term stable expression of fluorescently traced tumor cells

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